Iwabata Kazuki, Koshiyama Akiyo, Yamaguchi Taiki, Sugawara Hiroko, Hamada Fumika N, Namekawa Satoshi H, Ishii Satomi, Ishizaki Takashi, Chiku Hiroyuki, Nara Takayuki, Sakaguchi Kengo
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
Nucleic Acids Res. 2005 Oct 12;33(18):5809-18. doi: 10.1093/nar/gki883. Print 2005.
Lim15/Dmc1 is a meiosis specific RecA-like protein. Here we propose its participation in meiotic chromosome pairing-related events along with DNA topoisomerase II. Analysis of protein-protein interactions using in vitro binding assays provided evidence that Coprinus cinereus DNA topoisomerase II (CcTopII) specifically interacts with C.cinereus Lim15/Dmc1 (CcLim15). Co-immunoprecipitation experiments also indicated that the CcLim15 protein interacts with CcTopII in vivo. Furthermore, a significant proportion of CcLim15 and CcTopII could be shown to co-localize on chromosomes from the leptotene to the zygotene stage. Interestingly, CcLim15 can potently activate the relaxation/catenation activity of CcTopII in vitro, and CcTopII suppresses CcLim15-dependent strand transfer activity. On the other hand, while enhancement of CcLim15's DNA-dependent ATPase activity by CcTopII was found in vitro, the same enzyme activity of CcTopII was inhibited by adding CcLim15. The interaction of CcLim15 and CcTopII may facilitate pairing of homologous chromosomes.
Lim15/Dmc1是一种减数分裂特异性的类RecA蛋白。在此我们提出它与DNA拓扑异构酶II一起参与减数分裂染色体配对相关事件。利用体外结合试验对蛋白质-蛋白质相互作用进行分析,结果表明灰盖鬼伞DNA拓扑异构酶II(CcTopII)与灰盖鬼伞Lim15/Dmc1(CcLim15)特异性相互作用。免疫共沉淀实验也表明CcLim15蛋白在体内与CcTopII相互作用。此外,相当一部分的CcLim15和CcTopII在细线期到偶线期阶段可共定位于染色体上。有趣的是,CcLim15在体外可有效激活CcTopII的松弛/连环活动,而CcTopII可抑制CcLim15依赖的链转移活性。另一方面,虽然在体外发现CcTopII可增强CcLim15的DNA依赖的ATP酶活性,但加入CcLim15可抑制CcTopII的相同酶活性。CcLim15与CcTopII的相互作用可能有助于同源染色体配对。